Epigallocatechin gallate impels osteogenic differentiation of human BMSCs by targeting the METTL3/SATB2/Wnt/β-catenin axis

Bone marrow mesenchymal stem cells (BMSCs) are pluripotent cell, and its osteogenic differentiation disorder can lead to osteoporosis and seriously affect the quality of life. Epigallocatechin gallate (EGCG) is a catechin that has not been thoroughly studied on BMSCs. Flow cytometry was used for detecting cell surface antigens to identify BMSCs. Cell viability was measured using cell counting kit-8 (CCK8). Alkaline phosphatase (ALP) and alizarin red s (ARS) staining were employed for analyzing osteogenic differentiation. The expression of related genes was examined by western blot and quantitative real-time PCR (qRT-PCR). For the study of mechanism, ENCORI database, SRAMP database, methylated RNA immunoprecipitation (MeRIP), RNA immunoprecipitation (RIP) and mRNA stability assay were utilized to predict and validate the interactions between the proteins. EGCG enhanced cell viability and osteogenic differentiation of BMSCs. Moreover, special AT-rich sequence-binding protein 2 (SATB2) knockdown reversed the effect of EGCG in BMSCs. Mechanically, methyltransferase 3 (METTL3) mediated the N 6 -methyladenosine (m6A) methylation of SATB2. -Furthermore, EGCG-mediated promotion on BMSCs was weakened with the absence of METTL3. In addition, the deficiency of METTL3 and SATB2 could -abolish the effect of EGCG on Wnt/β-catenin pathway. EGCG induced osteogenic differentiation of BMSCs by affecting the METTL3/SATB2-mediated Wnt/β-catenin pathway.